The primary goal of our research program is directed towards an understanding of cell wall biosynthesis on the molecular level. The synthesis, assembly, and amplification of this structure is catalyzed by an integrated series of membrane-associated enzymes that are carefully regulated by the organism. Our research will contribute to an understanding of these enzymes, membrane events in peptidoglycan systhesis, membrane-wall interrelationships, and membrane teichoic acid biosynthesis. In addition, it is our goal to apply the results of our studies to the design of antibacterial agents that inhibit the assembly of cross-linked peptidoglycan. Five specific objectives will be undertaken for the coming year: (1) Intramolecular translocation of lipid intermediates in nascent peptidoglycan synthesis studied with fluorescent probes. (2) Factors involved in the assembly of the membrane-wall system for the incorporation of nascent peptidoglycan into wall. (3) Mechanism of penicilin and D-methionine action on nascent peptidoglycan synthesis. (4) Biosynthesis of membrane teichoic acid (D-ananyl-LTA): role of lypophilic intermediates studies with phosphodierterase II. (5) Purification and mechanistic analysis of phospho-MurNAc-pentapeptide translocase including lipid-translocase interacterations. An investigation of these projects will contribute to our understanding of cell wall assembly and membrane chemistry that relates to this process. Our present objectives will provide insights into the events, regulations, and translocations that occur in the bacterial membrane for elaboration of this extracellular structure, the cell wall.